Marine fungus secondary metabolite and preparation and application thereof
A technology of secondary metabolites, marine fungi, applied in the field of marine fungi, can solve the problems of no metabolites, research and application limitations, etc.
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Embodiment 1
[0048] Trichoderma harzianum ABC19819 and Penicillium brefeldianum ABC190807 were mixed and fermented, and the fermentation process was as follows:
[0049] (1) 200g of potatoes and 20g of glucose were mixed with sea water and the volume was adjusted to 1000mL to obtain a seed liquid culture medium;
[0050](2) Mix 80 g of rice and 100 mL of seawater to obtain a fermentation medium;
[0051] (3) activating the preserved two strains for subsequent use;
[0052] (4) Put the purified 2 strains of fungi into 5 bottles of seed liquid culture medium respectively, at 26°C, 180r min -1 Cultivate for 3 days under certain conditions as seed solution;
[0053] (5) Inoculate 3mL of the seed liquid of the two fungi into the fermentation medium, and carry out 135 bottles of fermentation culture based on 26°C static mixed fermentation culture for 45 days to obtain a fermented product.
Embodiment 2
[0055] The secondary product in the fermented product that embodiment 1 obtains is separated and extracted, and its process is:
[0056] (1) After the fermentation is over, sterilize and soak each fermentation bottle with ethyl acetate for 3 days, then filter and concentrate under reduced pressure to obtain the fermented product extract, repeat this process 3 times, and combine all the fermented product extracts , a total of 394.14g;
[0057] (2) Add 60-80 mesh silica gel with a mass ratio of 1:1 to the fermented product extract to mix the sample;
[0058] (3) Put the mixed sample after mixing into a silica gel column filled with 200-300 mesh silica gel:
[0059] (4) Through the petroleum ether-ethyl acetate system, the volume ratio is 100:0, 95:5, 90:10, 85:15, 80:20, 75:25, 70:30, 65:35, 60:40, 55:45, 50:50, 45:55, 40:60, 35:65, 20:80, 0:100 Carry out atmospheric pressure solvent elution on silica gel column chromatography in turn, each volume ratio, each eluent collected ...
Embodiment 3
[0123] Screen the insecticidal activity of the isolated metabolites as follows:
[0124] The resulting 6 compounds were prepared in 500 μg mL -1 The solution was screened for killing the 3rd instar larvae of Aedes aegypti by dipping method. And the same concentration of rotenone was prepared in the same way as the control group, and acetone was used as the blank control.
[0125] Transfer 10mL of prepared compound medicine solution or rotenone solution or acetone solution into disposable cups respectively, add 20 3rd instar Aedes aegypti larvae into each cup, repeat each treatment three times, and observe the records for 12, 24, 36, and 72 hours The number of dead Aedes aegypti larvae was counted, and the results were as follows:
[0126] The lethality of table 1 compound 1~6 to the 3rd instar larva of Aedes aegypti ( n=3)
[0127]
[0128] 1)Different lowercase letters after the data in the same column indicate significant difference (P<0.05, DMRT method)
[0129] ...
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